Synthesis and characterization of chitosan-based biomaterials modified with different active groups and their relationship with cytotoxicity

The cytotoxicity profile of three chitosan derivatives with different affinity to water was evaluated in vitro. The derivatives selected were carboxymethylated-chitosan (CMCH), linoleic acid modified-chitosan (LACH) and deoxycholic acid modified-chitosan (DACH), respectively, and the results of FTIR and NMR confirmed the successful modification. Cytotoxicity of these polymers was investigated via the red blood cell lysis assay and the MTT assay. The red blood cell lysis test showed that CH elicited a certain level of red blood cell toxicity, while CMCH, LACH and DACH all displayed low membrane damaging effects, with the hemolysis rates of 2.385%, 1.560% and 4.404%, respectively, which comes well within permissible limit (5%). The MTT assay revealed that CH exhibited significant inhibitory effect on fibroblast proliferation at higher concentration, while its three derivatives showed no cytotoxicity. CMCH had stimulatory effects on fetal mouse fibroblast proliferation. Differences in cytotoxicity of CH and its derivatives may result from the specific chemical modifications leading to the alteration of molecular charge density and type of the cationic functionalities, structure and sequence, and conformational flexibility.